Antibodies to the Aurora Kinase Family - FocusOn 123

Selected Antibody Panel to Aurora Kinase Subfamily

Acris Antibodies offers a collection of polyclonal and monoclonal antibodies to members of the Aurora kinase family. These antibodies are thoroughly evaluated and suitable for Western blotting and immunohistochemistry methods, and also for other immunological applications, such as immunocytochemistry and immunoprecipitation.

Introduction

Aurora kinases are serine/threonine kinases (STKs) that are essential for cell proliferation. Chromosomal segregation during mitosis as well as meiosis is regulated by kinases and phosphatases. The Aurora kinases associate with microtubules during chromosome movement and segregation. Aurora kinase B localizes to microtubules near kinetochores, specifically to the specialized microtubules called K-fibers, and Aurora kinase A localizes to centrosomes. Aurora kinases have generated significant interest in the cancer research field due to their elevated expression profiles in many human cancers. Three enzymes have been identified in mammalian cells to date:

Aurora kinase A (AURKA,STK6)

Zhou et al. (1998, PMID 9771714) found that Aurora kinase A is involved in the induction of centrosome duplication-distribution abnormalities and aneuploidy in mammalian cells. Centrosomes appear to maintain genomic stability through the establishment of bipolar spindles during cell division, ensuring equal segregation of replicated chromosomes to two daughter cells. Deregulated duplication and distribution of centrosomes are implicated in chromosome segregation abnormalities, leading to aneuploidy seen in many cancer cell types. In addition, amplification of STK6 is demonstrated in approximately 12 % of primary breast tumors, as well as in breast, ovarian, colon, prostate, neuroblastoma and cervical cancer cell lines. High expression of STK6 mRNA is detected in tumor cell lines without evidence of gene amplification. Ectopic expression of STK6 in mouse NIH 3T3 cells leads to the appearance of abnormal centrosome number (amplification) and transformation in vitro. Finally, overexpression of STK6 in near-diploid human breast epithelial cells reveals similar centrosome abnormality, as well as induction of aneuploidy. Aurora kinase A is a key regulatory component of the p53 pathway, and overexpression of Aurora kinase A leads to increased degradation of p53, causing downregulation of checkpoint-response pathways and facilitating oncogenic transformation of cells. These findings suggested that AURKA is a critical kinase-encoding gene, whose overexpression leads to centrosome amplification, chromosomal instability and transformation in mammalian cells.

Fig. 1: Human testis (formalin-fixed, paraffin-embedded; FFPE) stained with Acris Aurora kinase A antibody Cat.-No. AP07481PU-N,1:50, followed by biotinylated secondary antibody, alkaline phosphatase-streptavidin and chromogen
Fig. 1: Human testis (formalin-fixed, paraffin-embedded; FFPE) stained with Acris Aurora kinase A antibody Cat.-No. AP07481PU-N,1:50, followed by biotinylated secondary antibody, alkaline phosphatase-streptavidin and chromogen
Fig. 2: Detection of endogenous Aurora kinase A: Whole cell extracts of vanadate treated tumor cells (20,000 cells per lane) were applied to SDS-PAGE and transferred to a PVDF membrane. The immunoblot was probed with Acris Aurora kinase A antibody clone 7F12 Cat.-No. AM20207PU-N (0.5 µg/ml) for 1h at RT and developed by ECL: lane 1: A431; lane 2: SW480; lane 3: SW620; lane 4: HT29; lane 5: MCF-7; lane 6: MDA-MB231; lane 7: T47D
Fig. 2: Detection of endogenous Aurora kinase A: Whole cell extracts of vanadate treated tumor cells (20,000 cells per lane) were applied to SDS-PAGE and transferred to a PVDF membrane. The immunoblot was probed with Acris Aurora kinase A antibody clone 7F12 Cat.-No. AM20207PU-N (0.5 µg/ml) for 1h at RT and developed by ECL: lane 1: A431; lane 2: SW480; lane 3: SW620; lane 4: HT29; lane 5: MCF-7; lane 6: MDA-MB231; lane 7: T47D
Fig. 3: Immunoperoxidase staining of pancreatic cancer tissue (FFPE) with Acris Aurora kinase A antibody Cat.-No. SP2058P
Fig. 3: Immunoperoxidase staining of pancreatic cancer tissue (FFPE) with Acris Aurora kinase A antibody Cat.-No. SP2058P
Fig. 4: Immunofluorescence of monoclonal Abnova antibody to Aurora kinase A Cat.-No. H00006790-M01 on HeLa cell [antibody concentration 30 ug/ml]
Fig. 4: Immunofluorescence of monoclonal Abnova antibody to Aurora kinase A Cat.-No. H00006790-M01 on HeLa cell [antibody concentration 30 ug/ml]

Aurora Kinase B (AURKB, STK12)

This protein is a Ser/Thr protein kinase member of the Aurora subfamily that may be directly involved in regulating the cleavage of polar spindle microtubules, and is a key regulator for the onset of cytokinesis during mitosis. Aurora kinase B is localized to the midzone of the central spindle in late anaphase, concentrated into the midbody in telophase and cytokinesis, and is colocalized with gamma tubulin in the midbody. High levels of Aurora kinase B expression are seen in the thymus, although it is also expressed in the spleen, lung, testis, colon, placenta and fetal liver. Aurora kinase B is expressed during S and G2/M phase, and expression is up-regulated in cancer cells during M phase.

Fig. 5: Model of Aurora kinase B, inner centromere protein (INCENP) and survivin interactions and regulation. Survivin binds the N-terminus of INCENP, whereas the N terminus of Aurora kinase B binds the C-terminus of INCENP
Fig. 5: Model of Aurora kinase B, inner centromere protein (INCENP) and survivin interactions and regulation. Survivin binds the N-terminus of INCENP, whereas the N terminus of Aurora kinase B binds the C-terminus of INCENP
Fig. 6: Human hepatocarcinoma tissue (FFPE) reacted with Acris Aurora kinase B antibody Cat.-No. AP13522PU-N using peroxidase-conjugated secondary antibody followed by AEC staining
Fig. 6: Human hepatocarcinoma tissue (FFPE) reacted with Acris Aurora kinase B antibody Cat.-No. AP13522PU-N using peroxidase-conjugated secondary antibody followed by AEC staining

Aurora Kinase C (AURKC, STK13)

This protein acts as a chromosomal passenger protein and forms complexes with Aurora kinase B and inner centromere protein (INCENP). Aurora kinase C phosphorylates histone H3 in vitro. INCENP binds and activates AURKC in vivo and in vitro, and AURKC co-expressed with INCENP elicits histone H3 phosphorylation of interphase cells. Using an in vitro kinase assay, Yan et al. (2005, PMID 15938719) found that AURKC-SV phosphorylates a test substrate protein, and its activity depends on Thr179. During cell mitosis, AURKC-SV associates with chromosomes in prophase and metaphase, and then transferred to the central spindle midzone and the cortex where the contracting ring forms during the transition from anaphase to telophase. It remains in the midbody during cytokinesis.

Fig. 7: In the immunofluorescence experiment, staining of HeLa cells expressing GFP-Aurora kinase C is performed at different cellular mitotic stages with the Acris Aurora kinase C antibody Cat.-No. AP13519PU-N as primary antibody (column A), GFP fluorescence (column B), DAPI nuclear staining (column C), and anti-Aurora kinase C merged to DAPI staining (column D). Data is kindly provided by Drs. K. Sasai and S. Sen,University of Texas, MD Anderson Cancer Center (Houston, TX)
Fig. 7: In the immunofluorescence experiment, staining of HeLa cells expressing GFP-Aurora kinase C is performed at different cellular mitotic stages with the Acris Aurora kinase C antibody Cat.-No. AP13519PU-N as primary antibody (column A), GFP fluorescence (column B), DAPI nuclear staining (column C), and anti-Aurora kinase C merged to DAPI staining (column D). Data is kindly provided by Drs. K. Sasai and S. Sen,University of Texas, MD Anderson Cancer Center (Houston, TX)

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Primary Antibodies

Catalog No. Host Iso. Clone Pres. React. Applications  

Aurora-A/B (C-term) (incl. pos. control) antibody

Detection of endogenous Aurora-A/B: Whole cell extracts of vanadate treated tumor cells (20.000 cells per lane) were applied to SDS-PAGE and transferred to a PVDF membrane. The Immunoblot was probed with monoclonal Aurora-A/B antibody AM20208PU-N (5F11) (0.5 µg/ ml) for 1h at RT and developed by ECL (exp. time: 30 sec).
Lane 1: A431 
Lane 2: SW480 
Lane 3: SW620 
Lane 4: HT29
Lane 5: MCF-7 
Lane 6: MDA-MB231 
Lane 7: T47D Mouse IgG1 5F11 Purified Hu F, ICC/IF, WB
0.1 mg / €430.00
  Acris Antibodies GmbH

Aurora kinase A (N-term) (incl. pos. control) antibody

Detection of endogenous Aurora-A: Whole cell extracts of vanadate treated tumor cells (20.000 cells per lane) were applied to SDS-PAGE and transferred to a PVDF membrane. The Immunoblot was probed with monoclonal antibody AM20207PU-N (7F12) (0.5 µg/ ml) for 1h at RT and developed by ECL (exp. time: 30 sec).
Lane 1: A431 
Lane 2: SW480 
Lane 3: SW620 
Lane 4: HT29
Lane 5: MCF-7 
Lane 6: MDA-MB231 
Lane 7: T47D Mouse IgG1 7F12 Purified Can, Hu, Ms, Rt WB
0.1 mg / €430.00
  Acris Antibodies GmbH

Aurora kinase A pThr288 antibody

AP07481PU-N Aurora kinase A antibody staining of Formalin-Fixed Paraffin-Embedded Human Testis at 1/50 followed by biotinylated Goat anti-Rabbit  IgG secondary antibody, Alkaline Phosphatase-Streptavidin and Chromogen. Rabbit IgG Aff - Purified Hu P, WB
50 µl / €470.00
  Acris Antibodies GmbH

Aurora kinase A (C-term) antibody

Thymus: Formalin-Fixed Paraffin-Embedded (FFPE) Goat Aff - Purified Hu, Mky, Prim E, IP, P, WB
50 µg / €470.00
  Acris Antibodies GmbH

Aurora kinase A pThr288 antibody

Immunohistochemical analysis of paraffin-embedded human breast carcinoma tissue using Aurora A (phospho-Thr288) Antibody AP08036PU Rabbit Aff - Purified Hu, Ms P, WB
0.1 ml / €320.00
  Acris Antibodies GmbH

Aurora kinase A pThr288 antibody

Immunohistochemical analysis of paraffin-embedded human breast carcinoma tissue using Aurora A (phospho-Thr288) Antibody AP08036PU Rabbit Aff - Purified Hu, Ms P, WB
50 µl / €240.00
  Acris Antibodies GmbH

Aurora kinase A (N-term) antibody

Formalin-Fixed, Paraffin-Embedded Human cancer (Left) and Human colon carcinoma (Right) tissues  reacted with AP14756PU-N Aurora Kinase A antibody using peroxidase-conjugated secondary antibody followed by DAB staining. Rabbit Ig Purified Hu P, WB
0.4 ml / €370.00
  Acris Antibodies GmbH

Aurora kinase A antibody

Western blot analysis of recombinant Aurora-A kinase protein probed with SM2084B and F(ab)2 rabbit anti-mouse IgG:HRP (SP1012HRP). Mouse IgG2b 35C1 Biotin Hu, Ms ICC/IF, WB
0.1 mg / €450.00
  Acris Antibodies GmbH

Aurora kinase A antibody

Aurora kinase A Mouse IgG2b 35C1 Purified Hu, Ms ICC/IF, IP, WB
0.1 mg / €410.00
  Acris Antibodies GmbH

Aurora kinase A antibody

SP2058P Aurora kinase A antibody staining of Paraffin Embedded Human pancreatic cancer tissue sections using DAB staining detection. Rabbit Aff - Purified Hu IP, P
50 µg / €320.00
  Acris Antibodies GmbH

Aurora kinase A antibody

Immunofluorescence of monoclonal antibody to STK6 on HeLa cell. [antibody concentration 30 ug/ml] Mouse IgG2a 5F8 Purified Hu E, ICC/IF, WB
0.1 mg / €370.00
  Abnova Taiwan Corp.

Aurora kinase B antibody

Immunofluorescence of human HeLa cells stained with Hoechst 3342 (Blue) for chromatin staining and monoclonal anti-human Aurora kinase B antibody Cat.-No. AM09372PU (1:2000) with Texas Red (Red). Mouse IgG2b AT2B1 Purified Hu E, F, ICC/IF, WB
0.1 ml / €310.00
  Acris Antibodies GmbH

Aurora kinase B antibody

Immunofluorescence of human HeLa cells stained with Hoechst 3342 (Blue) for chromatin staining and monoclonal anti-human Aurora kinase B antibody Cat.-No. AM09372PU (1:2000) with Texas Red (Red). Mouse IgG2b AT2B1 Purified Hu E, F, ICC/IF, WB
50 µl / €230.00
  Acris Antibodies GmbH

Aurora kinase B (227-238) antibody

Spleen: Formalin-Fixed Paraffin-Embedded (FFPE) Rabbit Aff - Purified Hu E, ICC/IF, P, WB
50 µg / €470.00
  Acris Antibodies GmbH

Aurora kinase B (223-234) pThr232 antibody

Tonsil: Formalin-Fixed Paraffin-Embedded (FFPE) Rabbit IgG Aff - Purified Hu, Ms E, ICC/IF, P, WB
50 µg / €470.00
  Acris Antibodies GmbH

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